LOCUS       IL2_C125S_in_pQE        5115 bp    DNA     circular UNA 27-OCT-2017
DEFINITION  Concatenation of 2 sequences.
ACCESSION   urn.local...1l-88474yn
VERSION     urn.local...1l-88474yn
KEYWORDS    .
SOURCE      
  ORGANISM  
            .
FEATURES             Location/Qualifiers
     misc_feature    1..410
                     /Source="referenceToDocument: urn:local:.:18-8847342"
                     /label="Digestion of IL2 C125S - Fragment 2 (concatenated
                     sequence 1)"
                     /note="Geneious type: Concatenated sequence"
     ligation        1..4
                     /label="Ligation"
     misc_feature    379..380
                     /Original_Bases="GT"
                     /label="GT"
                     /note="Geneious type: Editing History Replacement"
     misc_feature    404^405
                     /Original_Bases="GGTGGAGGAGGTTCTGGAGGCGGTGGAAGTGGTGGCGGAGG
                     TAGCGCTTGTGACGACCAAAATCCCTGGCGTGCTTACCTGGATCTCCTCTTCCCTACC
                     GACACACTGCTGCTGGATCTCCTGTGG"
                     /label="GGTGGAGGAGGTTCTGGAGGCGGTGGAAGTGGTGGCGGAGGTAGCGCTTG
                     TGACGACCAAAATCCCTGGCGTGCTTACCTGGATCTCCTCTTCCCTACCGACACACTG
                     CTGCTGGATCTCCTGTGG"
                     /note="Geneious type: Editing History Deletion"
     ligation        407..410
                     /label="Ligation"
     terminator      427..521
                     /note="lambda t0 terminator"
                     /note="transcription terminator from phage lambda"
                     /label="lambda t0 terminator"
     CDS             565..1224
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /note="CmR"
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAF
                     LKTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETF
                     SSLWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNV
                     ANMDNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGG
                     A"
                     /label="cat CDS"
     terminator      1289..1375
                     /gene="Escherichia coli rrnB"
                     /note="rrnB T1 terminator"
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /label="Escherichia coli rrnB terminator"
     CDS             complement(1468..2550)
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAE
                     LNYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERS
                     GVEACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSII
                     FSHEDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREG
                     DWSAMSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDT
                     EDSSCYIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNT
                     QTASPRALADSLMQLARQVSRLESGQ"
                     /label="lacI CDS"
     promoter        complement(2551..2628)
                     /gene="lacI (mutant)"
                     /note="lacIq promoter"
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about
                     10-fold."
                     /label="lacI (mutant) promoter"
     misc_feature    2820..2960
                     /note="bom"
                     /note="basis of mobility region from pBR322"
                     /label="bom"
     rep_origin      complement(3146..3734)
                     /direction="LEFT"
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /label="ori rep origin"
     CDS             complement(3905..4765)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVE
                     YSPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRL
                     DRWEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPL
                     LRSALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIA
                     EIGASLIKHW"
                     /label="bla CDS"
     promoter        complement(4766..4870)
                     /gene="bla"
                     /note="AmpR promoter"
                     /label="bla promoter"
     origin          4970^4971
                     /label="origin"
     promoter        4980..5024
                     /note="T5 promoter"
                     /note="bacteriophage T5 promoter for E. coli RNA 
                     polymerase, with embedded lac operator"
                     /label="T5 promoter"
     protein_bind    5000..5016
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator protein bind"
     protein_bind    5032..5048
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be
                     relieved by adding lactose or
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator protein bind"
     primer_bind     5039..5058
                     /Mismatches=0
                     /created_by="primer3"
                     /annotation_group="2607104510194d9f7505f-2d12-4510-9c20-d0
                     bf142d16ec"
                     /%GC="40.0"
                     /Tm="52.6"
                     /Hairpin_Tm="None"
                     /Self_Dimer_Tm="None"
                     /Pair_Dimer_Tm="None"
                     /Sequence="CGGATAACAATTTCACACAG"
                     /Product_Size=175
                     /label="pQE80L fwd"
     RBS             5067..5078
                     /note="strong bacterial ribosome binding site (Elowitz and
                      Leibler, 2000)"
                     /label="RBS"
     CDS             5097..5114
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
                     /label="6xHis affinity tag CDS"
ORIGIN      
        1 gatccgcacc tacttcaagt tctacaaaga aaacacagct acaactggag catttactgc
       61 tggatttaca gatgattttg aatggaatta ataattacaa gaatcccaaa ctcaccagga
      121 tgctcacatt taagttttac atgcccaaga aggccacaga actgaaacat cttcagtgtc
      181 tagaagaaga actcaaacct ctggaggaag tgctaaattt agctcaaagc aaaaactttc
      241 acttaagacc cagggactta atcagcaata tcaacgtaat agttctggaa ctaaagggat
      301 ctgaaacaac attcatgtgt gaatatgctg atgagacagc aaccattgta gaatttctga
      361 acagatggat taccttttcc caaagcatca tctcaacact gacttaagct taattagctg
      421 agcttggact cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca
      481 gaacgctcgg ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga
      541 ttttcaggag ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat
      601 atatcccaat ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc
      661 tataaccaga ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag
      721 cacaagtttt atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa
      781 tttcgtatgg caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac
      841 accgttttcc atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat
      901 ttccggcagt ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc
      961 tatttcccta aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt
     1021 ttcaccagtt ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc
     1081 atgggcaaat attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat
     1141 catgccgttt gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc
     1201 gatgagtggc agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct
     1261 ggggtaatga ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg
     1321 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc
     1381 ctctagatta cgtgcagtcg atgataagct gtcaaacatg agaattgtgc ctaatgagtg
     1441 agctaactta cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     1501 tgccagctgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     1561 cagggtggtt tttcttttca ccagtgagac gggcaacagc tgattgccct tcaccgcctg
     1621 gccctgagag agttgcagca agcggtccac gctggtttgc cccagcaggc gaaaatcctg
     1681 tttgatggtg gttaacggcg ggatataaca tgagctgtct tcggtatcgt cgtatcccac
     1741 taccgagata tccgcaccaa cgcgcagccc ggactcggta atggcgcgca ttgcgcccag
     1801 cgccatctga tcgttggcaa ccagcatcgc agtgggaacg atgccctcat tcagcatttg
     1861 catggtttgt tgaaaaccgg acatggcact ccagtcgcct tcccgttccg ctatcggctg
     1921 aatttgattg cgagtgagat atttatgcca gccagccaga cgcagacgcg ccgagacaga
     1981 acttaatggg cccgctaaca gcgcgatttg ctggtgaccc aatgcgacca gatgctccac
     2041 gcccagtcgc gtaccgtctt catgggagaa aataatactg ttgatgggtg tctggtcaga
     2101 gacatcaaga aataacgccg gaacattagt gcaggcagct tccacagcaa tggcatcctg
     2161 gtcatccagc ggatagttaa tgatcagccc actgacgcgt tgcgcgagaa gattgtgcac
     2221 cgccgcttta caggcttcga cgccgcttcg ttctaccatc gacaccacca cgctggcacc
     2281 cagttgatcg gcgcgagatt taatcgccgc gacaatttgc gacggcgcgt gcagggccag
     2341 actggaggtg gcaacgccaa tcagcaacga ctgtttgccc gccagttgtt gtgccacgcg
     2401 gttgggaatg taattcagct ccgccatcgc cgcttccact ttttcccgcg ttttcgcaga
     2461 aacgtggctg gcctggttca ccacgcggga aacggtctga taagagacac cggcatactc
     2521 tgcgacatcg tataacgtta ctggtttcac attcaccacc ctgaattgac tctcttccgg
     2581 gcgctatcat gccataccgc gaaaggtttt gcaccattcg atggtgtcgg aatttcgggc
     2641 agcgttgggt cctggccacg ggtgcgcatg atctagagct gcctcgcgcg tttcggtgat
     2701 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     2761 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     2821 gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat
     2881 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     2941 ggagaaaata ccgcatcagg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3001 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3061 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3121 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3181 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3241 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3301 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3361 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3421 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3481 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3541 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaaggaca gtatttggta
     3601 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     3661 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     3721 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     3781 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     3841 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     3901 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     3961 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4021 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4081 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4141 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4201 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4261 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4321 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4381 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4441 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4501 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4561 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4621 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     4681 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     4741 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     4801 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     4861 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca
     4921 tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtcttcac ctcgagaaat
     4981 cataaaaaat ttatttgctt tgtgagcgga taacaattat aatagattca attgtgagcg
     5041 gataacaatt tcacacagaa ttcattaaag aggagaaatt aactatgaga ggatcgcatc
     5101 accatcacca tcacg
//
